Monthly Archives: October 2011

Monday 10/31/11

Posted on October 31, 2011 by admin

10:00 A — 11:00 P Colenso’s lab meeting PureLink DNA extraction — visible pellet but no DNA detected on nano-drop.  Poor resupension?  tried vortex and extended incubation at 37C.  No improvement in conc. Test digest, try to detect by gel? … Continue reading →

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Sunday 10/30/11

Posted on October 30, 2011 by admin

8:00 P – 11:00 P Finish washing out secondary antibodies from mixed mRNA/protein in situs. Add Hoechst stain to all embryos Mount half of embryos directly for confocal imaging, move other half to ethanol in prep for resin embedding. Skipped … Continue reading →

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Saturday 10/29/11

Posted on October 29, 2011 by admin

  wash out primary antibodies on mixed protein/mRNA staining in situs.

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Wednesday 10/26/11

Posted on October 26, 2011 by admin

10:00 A -7:00 P, 10:00P – 12:00A Start PBS rinses of primary antibody for standard in situs Change antibody embedding samples to 70% resin Start hybe washes for new chamber in situs — be sure that filter does not slip … Continue reading →

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Tuesday 10/25/11

Posted on October 25, 2011 by admin

10:00A – 8:40 P, 9:45P – 12:00A in situs day 2 yw in situ in new embryo chamber, day 1 move antibodies to opaque box in 4C under Storm2/Graham bench. use on-deck 2 mL tubes to keep hot-wash solution warm … Continue reading →

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complete fluid exchange embryo chambers 3.0

Posted on October 25, 2011 by admin

To build: cut a small gasket 3-5mm from thin plastic tubing, diameter equal to eppendorf. Fit this snuggly around a P1000 pipette tip so the resulting plastic and rubber-gasket fits very tightly in an eppendorf. Remove the inner plastic, put … Continue reading →

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Antibodies from White study

Posted on October 25, 2011 by admin

Commercially purchased: H3K9Ac ab4441 Abcam H3K9me3 ab8898 Abcam RNA Pol II 8wG16 Covance H3K4me1 ab8895 Abcam H3K4me3 ab8580 Abcam H3K27Ac ab4729 Abcam H3K27me3 07-449 Upstate H3K36me3 ab9050 Abcam HP1 ab25726 Abcam HP1 Covance a-H3K4me3-LP LP Bio a-end300 Santa Cruz

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Monday 10/24/11

Posted on October 24, 2011 by admin

9:45 A – 10:45 P Embryos for in situ into hybe.  New hybe soln in door in -20C freezer. wash out primary antibody for antibody stains flip fly stocks make new top and side access vacuum embryo chambers snaD line … Continue reading →

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Sunday 10/23/11

Posted on October 23, 2011 by admin

9:30 A — 11:00A, 3:30P – 7:30P, 8:50P – 11:30P discard failed restriction digest, nicked translation, etc.  Wait for new columns to repeat DNA extraction. Start new stain of yw for bcd, hb, and laminin in situs day 1 — … Continue reading →

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Saturday 10/22/11

Posted on October 22, 2011 by admin

3:00P – 7:00P, 11:00P – 12:00A test gel for repeat trial of nicked translation — still no DNA detectable on gel, despite DNA detectable on spec. Trying straight gel of original isolated DNA — still no band test PCR of … Continue reading →

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